PKHhigh/CD133+/CD24− Renal Stem-Like Cells Isolated from Human Nephrospheres Exhibit In Vitro Multipotency

Abstract

The mechanism upon which human kidneys undergo regeneration is debated, though different lineage-tracing mouse models have tried to explain the cellular types and the mechanisms involved. Different sources of human renal progenitors have been proposed, but it is difficult to argue whether these populations have the same capacities that have been described in mice. Using the nephrosphere (NS) model, we isolated the quiescent population of adult human renal stem-like PKHhigh/CD133+/CD24− cells (RSC). The aim of this study was to deepen the RSC in vitro multipotency capacity. RSC, not expressing endothelial markers, generated secondary nephrospheres containing CD31+/vWf+ cells and cytokeratin positive cells, indicating the coexistence of endothelial and epithelial commitment. RSC cultured on decellularized human renal scaffolds generated endothelial structures together with the proximal and distal tubular structures. CD31+ endothelial committed progenitors sorted from nephrospheres generated spheroids with endothelial-like sprouts in Matrigel. We also demonstrated the double commitment toward endothelial and epithelial lineages of single RSC. The ability of the plastic RSC population to recapitulate the development of tubular epithelial and endothelial renal lineages makes these cells a good tool for the creation of organoids with translational relevance for studying the parenchymal and endothelial cell interactions and developing new therapeutic strategies.

The therapeutic potential of an allosteric non‐competitive CXCR1/2 antagonist for diabetic nephropathy

“…The first point of innovation of the present study lies in the use of a human NS model to investigate the role of the IL‐8‐CXCR1/2 axis. This model 15,35 represents in fact a unique tool and a human source of primary renal cells, with emphasis on podocytes, which are normally difficult to isolate and can be cultured only upon immortalisation 36 . In a previous study, Loretelli et al.…”

“…These results suggest a differential involvement of epithelial and podocyte cells in DN pathogenesis, which leads to a differential Ladarixin effect on epithelial cells, podocytes and leucocytes contribution to disease progression (Figure 5). This model 15,35 represents in fact a unique tool and a human source of primary renal cells, with emphasis on podocytes, which are normally difficult to isolate and can be cultured only upon immortalisation. 36 In a previous study, Loretelli et al showed that in vitro challenge of human immortalised podocytes with HG resulted in increased IL-8 secretion and DNA damage, and that this damage is prevented by CXCR1/2 blockade.…”

Role of extracellular matrix components and structure in new renal models in vitro

“…These scaffolds allow for the investigation of cell-ECM interactions in specific organ or tissue microdomains. For example, the fate of pluripotent or progenitor cells in a recellularized scaffold can be followed to learn about specific cell differentiation determinants ( Du et al, 2016 ; Bombelli et al, 2018 ; Zhang et al, 2019 ; Bombelli et al, 2020 ; Ullah et al, 2020 ).…”